Contents

Overview

This is the protocol used by P. Xu for fixing yeast cells for GFP fluorescence.

Materials

Yeast cells

Formaldehyde 37% (Sigma-Aldrich, #252549)

Phosphate buffered saline??

Protocol

Prepare eppendorf tubes with 50μL of 37% formaldehyde, one per sample.

Add 450μL of culture to 50μL of formaldehyde in an eppendorf tube and mix by inversion. (The goal is to have a final concentration of formaldehyde around 3.7%. So you can adjust the cell and formaldehyde volumes accordingly, as long as you end up with 3.7% formaldehyde).

Incubate the tube at room temperature for 15-20 minutes.

Spin down at 8000rpm for 1 minute.

Wash cells with 1X PBS 3 times, spin at 8000rpm for 1 minute each time (can be longer if you need).

Resuspend cell pellet in 100ul of 1X PBS.

Store samples at 4°C until you are ready to image.

Notes

Please feel free to post comments, questions, or improvements to this protocol.
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